High-Throughput Analysis of Serum Antigens Using Sandwich ELISAs on Microarrays

Servoss, Shannon L.; Gonzalez, Rachel; Varnum, Susan M.; Zangar, Richard C.

doi:10.1007/978-1-60327-811-9_10

Cited by 20 publications

(16 citation statements)

References 12 publications

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“…Slides were printed using a piezoelectric GeSiM NanoPlotter 2 and processed as described previously (Servoss et al, 2009). All ELISA reagents for the 18 cytokine assays (amphiregulin (AMR), eotaxin, granulocyte-colony stimulating factor (GCSF), interleukins 1α, 6, and 12 (IL-1α, 6, and 12), GRO1 oncogene (KC; CXCL1), matrix metalloproteinase 2 (MMP2), small inducible cytokine A5 (RANTES; CCL5), thymus and activation-regulated chemokine (TARC; CCL17), tumor necrosis factor (TNFα), vascular endothelial growth factor A (VEGF), monocyte chemotactic protein 1 (MCP1), macrophage-derived cytokine (MDC), macrophage inflammatory proteins 1α (MIP-1α, CCL3), 1β (MIP-1β,CCL4), 1γ (MIP-1γ,CCL9) and 2 (MIP-2, CXCL2)) were purchased from R&D Systems.…”

Section: Methodsmentioning

confidence: 99%

Susceptibility to quantum dot induced lung inflammation differs widely among the Collaborative Cross founder mouse strains

Scoville

White

Botta

et al. 2015

Toxicology and Applied Pharmacology

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Quantum dots (QDs) are engineered semiconductor nanoparticles with unique physicochemical properties that make them potentially useful in clinical, research and industrial settings. However, a growing body of evidence indicates that like other engineered nanomaterials, QDs have the potential to be respiratory hazards, especially in the context of the manufacture of QDs and products containing them, as well as exposures to consumers using these products. The overall goal of this study was to investigate the role of mouse strain in determining susceptibility to QD-induced pulmonary inflammation and toxicity. Male mice from 8 genetically diverse inbred strains (the Collaborative Cross founder strains) were exposed to CdSe–ZnS core–shell QDs stabilized with an amphiphilic polymer. QD treatment resulted in significant increases in the percentage of neutrophils and levels of cytokines present in bronchoalveolar lavage fluid (BALF) obtained from NOD/ShiLtJ and NZO/HlLtJ mice relative to their saline (Sal) treated controls. Cadmium measurements in lung tissue indicated strain-dependent differences in disposition of QDs in the lung. Total glutathione levels in lung tissue were significantly correlated with percent neutrophils in BALF as well as with lung tissue Cd levels. Our findings indicate that QD-induced acute lung inflammation is mouse strain dependent, that it is heritable, and that the choice of mouse strain is an important consideration in planning QD toxicity studies. These data also suggest that formal genetic analyses using additional strains or recombinant inbred strains from these mice could be useful for discovering potential QD-induced inflammation susceptibility loci.

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Section: Methodsmentioning

confidence: 99%

Susceptibility to quantum dot induced lung inflammation differs widely among the Collaborative Cross founder mouse strains

Scoville

White

Botta

et al. 2015

Toxicology and Applied Pharmacology

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“…Quantitative, multiplexed, sandwich ELISA microarray analysis was undertaken as described previously [17, 18]. We have previously demonstrated that the multiplexed assays used in the current study do not have any detectable cross-assay interference, and that this analytical platform can quantitatively detect purified antigens spiked into human sera [17].…”

Section: Methodsmentioning

confidence: 99%

Reactive oxygen species alter autocrine and paracrine signaling

Zangar

Bollinger

Weber

et al. 2011

Free Radical Biology and Medicine

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Cytochrome P450 (P450) 3A4 (CYP3A4) is the most abundant P450 protein in human liver and intestine, and is highly inducible by a variety of drugs and other compounds. The P450 catalytic cycle is known to uncouple and release reactive oxygen species (ROS), but the effects of ROS from P450 and other enzymes in the endoplasmic reticulum has been poorly studied from the perspective of effects on cell biology. In this study, we express low levels of CYP3A4 in HepG2 cells, a human hepatocarcinoma cell line, and examine effects on intracellular levels of ROS and on the secretion of a variety of growth factors that are important in extracellular communication. Using the redox sensitive dye, RedoxSensor Red, we demonstrate that CYP3A4 expression increases levels of ROS in viable cells. A custom ELISA microarray platform was employed to demonstrate that expression of CYP3A4 increased secretion of amphiregulin, intracellular adhesion molecule 1, matrix metalloprotease 2, platelet-derived growth factor (PDGF) and vascular endothelial growth factor but suppressed secretion of CD14. The antioxidant, N-acetylcysteine, suppressed all P450-dependent changes in protein secretion except for CD14. Quantitative RT-PCR demonstrated that changes in protein secretion were consistently associated with corresponding changes in gene expression. Inhibition of the NF-κB pathway blocked P450 effects on PDGF secretion. CYP3A4 expression also altered protein secretion in human mammary epithelial cells and C10 mouse lung cells. Overall, these results suggest that increased ROS production in the endoplasmic reticulum alters the secretion of proteins that have key roles in paracrine and autocrine signaling.

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“…Cell extracts containing the antigens were incubated on the chip and captured by the respective spotted antibodies. This step was followed by addition of labelled purified monoclonal antibodies recognising a different specific epitope of the antigen [59]. Avoidance of the labelling operation and achievement of specific signals were two important advantages associated with this assay.…”

Section: Antibody Microarray Assaysmentioning

confidence: 99%

Production, Novel Assay Development and Clinical Applications of Monoclonal Antibodies

Chiarella

2011

PRA

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Since the advent of hybridoma technology 35 years ago, research on monoclonal antibodies has developed enormously. Monoclonal antibodies of mouse origin were the first to be produced and continue to be the most popular affinity reagents for investigating the proteome of all organisms. For their adaptability to a variety of biological assays monoclonal antibodies are key tools for basic research as well as for diagnosis and therapy of human diseases. Recently, the expanding demand of high-quality antibodies with better specificities has resulted in a significant improvement in traditional hybridoma production methods. Owing to the ability of these affinity reagents to selectively target tumour cells, cancer has been a major focus of programmes for monoclonal antibody development. This review focuses on patents related to the advances made in the monoclonal antibody manufacture, showing how the traditional production techniques were turned into alternative, faster and more effective methods. Other patents are focussed on new technologies in which monoclonal antibodies are employed for the development of high-performance screening assays. A conclusive series of patents is related to monoclonal antibodies which find application to the diagnosis and the treatment of specific cancer diseases such as haematological malignancies and solid tumours.

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High-Throughput Analysis of Serum Antigens Using Sandwich ELISAs on Microarrays

Cited by 20 publications

References 12 publications

Susceptibility to quantum dot induced lung inflammation differs widely among the Collaborative Cross founder mouse strains

Susceptibility to quantum dot induced lung inflammation differs widely among the Collaborative Cross founder mouse strains

Reactive oxygen species alter autocrine and paracrine signaling

Production, Novel Assay Development and Clinical Applications of Monoclonal Antibodies

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